In order to determine the rate of evolution of the rGII.3, GII.3, GII.4 and GII.7 capsids, the nucleotide sequences of ORF2 were analysed. RNA capsid sequences used for the analysis included eight from this study and 76 sequences from GenBank, with the oldest strains available dating back to 1987. The strains used and their GenBank accession numbers are listed in Text S1. The rate of evolution (substitutions/nucleotide site/year) for GII.3, GII.b/GII.3 GII.4 and GII.7 NoVs was determined by calculating the number of nucleotide substitutions in ORF2 compared to an ancestral strain and this was plotted against time [27]. The rate of evolution was determined by linear regression with the program GraphPad PRISM version 4 and was equivalent to the gradient of the line. Pairwise alignments of RNA sequences and evolutionary distances between sequences were carried out using the Maximum Composite Likelihood model in Mega 4.0 [28]. Bootstrapped trees (1000 data sets) were constructed using the Neighbour-joining method, also with the program Mega 4.0.
Molecular Evolutionary Genetics Nei 1987 Pdf 291
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